CN110198740B - Novel BSH complexes for boron neutron capture therapy - Google Patents
Novel BSH complexes for boron neutron capture therapy Download PDFInfo
- Publication number
- CN110198740B CN110198740B CN201780084294.2A CN201780084294A CN110198740B CN 110198740 B CN110198740 B CN 110198740B CN 201780084294 A CN201780084294 A CN 201780084294A CN 110198740 B CN110198740 B CN 110198740B
- Authority
- CN
- China
- Prior art keywords
- bsh
- complex
- cancer
- peptide
- cells
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/095—Sulfur, selenium, or tellurium compounds, e.g. thiols
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/16—Fluorine compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/22—Boron compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/08—Peptides having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/009—Neutron capture therapy, e.g. using uranium or non-boron material
- A61K41/0095—Boron neutron capture therapy, i.e. BNCT, e.g. using boronated porphyrins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
- A61K47/6941—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a granulate or an agglomerate
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Inorganic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Dermatology (AREA)
- Molecular Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention provides: a complex of mercaptoundehydroceledodecaborate (BSH) and a peptide for use in Boron Neutron Capture Therapy (BNCT); a method of making the composite; and cancer therapy using the complex.
Description
Technical Field
The invention relates to a medicament for treating cancer and a preparation method thereof. In particular, the present invention relates to a complex of mercaptoundecahydridododecaborate (BSH) and a peptide for Boron Neutron Capture Therapy (BNCT), a method of preparing the same, and cancer therapy using the same.
Background
Boron Neutron Capture Therapy (BNCT) is excellentA method of treating cancer in which a boron isotope is used10B is introduced into cancer cells and irradiated with a neutron beam so that only cancer cells are killed, and QOL after treatment is also excellent. Safe and easy delivery of sufficient amounts of boron drugs into cancer cells is the biggest challenge of BNCT. BPA, which is an amino acid derivative containing one boron atom, has been used as a main agent, and BSH, which does not enter cells themselves including cancer cells, has been used as an auxiliary agent (see non-patent documents 1 and 2).
BPA enters cells through amino acid transporters present in the cell membrane. However, since even normal cells, in the case where they are actively expanding cells such as mucosal epithelial cells and hair growth cells, actively take in BPA, neutron irradiation damages these normal cells. Furthermore, since such amino acid transporters are "exchange transporters" (see non-patent document 3), BPA (LAT1 is considered to be the main responsible) entering cells through the amino acid transporters is not easily retained in large amounts in cells (cancer cells), and further BPA has only one boron atom per molecule, which results in poor collision efficiency for neutron beams, and thus large administration (several tens of grams per adult) is required to deliver and retain boron necessary in cancer cells.
BSH is a crystal with 12 boron atoms per molecule and therefore has high efficiency. However, BSH does not penetrate the cell membrane and therefore does not itself enter the cell. BSH leaks out of the delicate blood vessels of cancer tissues, enters interstitial fluid, and is only retained around cells. Therefore, secondary particles (α particles, Li nuclei) generated by neutron irradiation hardly reach the nuclei where gene DNA is affected, and thus a sufficient killing effect cannot be obtained. Further, the lack of cancer cell specificity of BSH is problematic.
In addition to boron drugs, BPA and BSH, which have been clinically used so far, various nanocarriers are proposed at present. They are classified into liposomes (lipid bilayer vesicles), polymeric micelles (amphiphilic polymer vesicles), and carbon nanotubes, but have the following problems. Liposomes release drug continuously and are physically unstable at low toxicity. Biocompatibility and biodegradability of the polymeric micelle are preferable, but the half-life in vivo is short. Carbon nanotubes have a large surface area but are not cargo selective and are toxic.
Documents of the prior art
Non-patent document
Non-patent document 1: kato i. et al, appl.radiat.isot.2004; 61:1069-73
Non-patent document 2: hatanaka h. et al, j.neurol.1975; 209:81-94
Non-patent document 3: biochemistry, Vol.86, No. 3, pp 338-
Disclosure of Invention
Problems to be solved by the invention
The problem addressed by the present invention is to provide a drug that can deliver BSH directly and retain it in cancer cells so that BNCT can be performed efficiently.
Means for solving the problems
The present inventors have conducted extensive studies to solve the above-mentioned problems, and have found that a complex including a peptide containing hydrophobic amino acid residues and basic amino acid residues and BSH obtained by mixing the peptide and BSH in an aqueous solution has a spherical shape with a diameter of about 20nm to about 200nm, which is an ideal shape for cell introduction, and is directly delivered to and retained in cancer cells, thereby completing the present invention.
Specifically, the following are provided by the present invention:
(1) a method of making a composite body comprising a peptide comprising a hydrophobic amino acid residue and a basic amino acid residue and mercaptoundecahydridodecaborate (BSH), the method comprising mixing the peptide and BSH in an aqueous solution;
(2) the method according to (1), wherein BSH is mixed in a ratio of 1mol to 1000mol with respect to 1mol of the peptide;
(3) the method of (1) or (2), further comprising adjusting the diameter of the complex;
(4) the method according to any one of (1) to (3), wherein the complex is spherical with a diameter of about 20nm to about 200 nm;
(5) the method according to any one of (1) to (4), wherein the peptide is represented by the following formula (1):
(X)m-(Z)n (1)
wherein each of the m amino acid residues X is independently alanine, valine, leucine, or glycine; n amino acid residues Z are each independently-NHCH (COOH) R1；R1Is- (CH)2)pNHR2；R2is-H or-C (NH) NH2(ii) a m is 4 to 10; n is 1 to 2; and p is 1 to 6;
(6) the method of (5), wherein X is alanine; m is 6; z is lysine, arginine, homoarginine, ornithine, 2, 7-diaminoheptanoic acid, 2, 4-diaminobutyric acid, or 2-amino-4-guanidinobutyric acid; and n is 1;
(7) the method of (6), wherein X is alanine; m is 6; z is lysine or arginine; and n is 1;
(8) a complex comprising a peptide comprising a hydrophobic amino acid residue and a basic amino acid residue and BSH;
(9) the composite of (8), wherein the composite is spherical with a diameter of about 20nm to about 200 nm;
(10) the complex according to (8) or (9), wherein the peptide is represented by the following formula (1):
(X)m-(Z)n (1)
wherein each of the m amino acid residues X is independently alanine, valine, leucine, or glycine; n amino acid residues Z are each independently-NHCH (COOH) R1；R1Is- (CH)2)pNHR2；R2is-H or-C (NH) NH2(ii) a m is 4 to 10; n is 1 to 2; and p is 1 to 6;
(11) the complex according to (10), wherein X is alanine; m is 6; z is lysine, arginine, homoarginine, ornithine, 2, 7-diaminoheptanoic acid, 2, 4-diaminobutyric acid, or 2-amino-4-guanidinobutyric acid; and n is 1;
(12) the complex according to (11), wherein X is alanine; m is 6; z is lysine or arginine; and n is 1;
(13) a medicament for boron neutron capture therapy of cancer, comprising the complex according to any one of (8) to (12);
(14) a method of treating cancer, comprising administering the complex according to any one of (8) to (12) to a cancer patient and irradiating the cancer patient with a neutron beam;
(15) the complex according to any one of (8) to (12), which is used for boron neutron capture therapy of cancer; and
(16) use of the complex according to any one of (8) to (12) in the manufacture of a medicament for boron neutron capture therapy of cancer.
ADVANTAGEOUS EFFECTS OF INVENTION
According to the present invention, a complex capable of delivering and retaining a large amount of BSH in cancer cells can be obtained by a simple operation such as mixing a peptide containing hydrophobic amino acid residues and basic amino acid residues with BSH in an aqueous solution. The ratio of this peptide to BSH is desirably in the range of an equivalent to 1000-fold molar excess of BSH. The effect of BNCT is significantly enhanced when using the composites of the invention. For example, BSH can indeed be delivered and retained in cancer cells by a single injection.
Drawings
FIG. 1 is a scanning electron micrograph showing A6K (trifluoroacetate salt, hereinafter TFA salt) in aqueous solution (concentration 50. mu.M). The bar (bar) at the lower right corner is 2 μm.
FIG. 2 is a graph showing the results of a Dynamic Light Scanning (DLS) test of A6K (TFA salt) (concentration 200. mu.M) in aqueous solution.
FIG. 3 is a scanning electron micrograph of a complex obtained by mixing A6K (TFA salt) (10. mu.M) with BSH (1000. mu.M) in an aqueous solution. The bar in the lower right corner is 500 nm.
FIG. 4 is a graph showing the results of DLS test of a complex obtained by mixing A6K (TFA salt) (200. mu.M) and BSH (2000. mu.M) in an aqueous solution.
Figure 5 is a graph showing the effect of mixing time of A6K (TFA salt) and BSH and culture time of complex with U87 Δ EGFR cells on the amount of complex delivered into the cells. The effect was studied using Inductively Coupled Plasma Atomic Emission spectrometry (ICP-AES). P < 0.005.
FIG. 6 is a scanning electron micrograph of a complex obtained by mixing A6R (TFA salt) (10. mu.M) and BSH (1000. mu.M) in an aqueous solution. The tape at the lower right corner was 1 micron.
FIG. 7 is a graph showing concentration-dependent cell introduction of a complex obtained by mixing A6R (TFA salt) and BSH in an aqueous solution. Inductively coupled plasma atomic emission spectrometry (ICP-AES) was used to study the amount of complex delivered into the cells. P <0.01, and p < 0.001.
Fig. 8 is a graph showing the effect of the complex obtained by mixing A6R (TFA salt) and BSH in aqueous solution and the culture time of U87 Δ EGFR cells on the amount of complex delivered into the cells. The effect was studied using inductively coupled plasma atomic emission spectrometry (ICP-AES).
Fig. 9 is a diagram showing U87 Δ EGFR intracellular distribution (subellular distribution) of BSH contained in a complex obtained by mixing A6K (TFA salt) and BSH in an aqueous solution using a specific antibody recognizing BSH.
FIG. 10 is a scanning electron micrograph of a complex obtained by mixing A6K (hydrochloride salt) (166. mu.M) and BSH (1.66mM) in an aqueous solution. The tape at the lower right corner was 500 nm.
FIG. 11 is a view showing cell introduction of a complex obtained by mixing A6K (hydrochloride salt) and BSH in an aqueous solution. Inductively coupled plasma atomic emission spectrometry (ICP-AES) was used to study the amount of complex delivered into the cells.
Fig. 12 is a graph showing the U87 Δ EGFR intracellular distribution of BSH contained in a complex obtained by mixing A6K (hydrochloride salt) and BSH in an aqueous solution using a specific antibody recognizing BSH. The left panel is a nuclear staining pattern showing the presence of cells. The middle panel is the local presence of BSH as shown by anti-BSH antibodies. The staining pattern and the right column were obtained by overlaying the left column on the middle column. The numbers on the left side of the figure are the A6K (hydrochloride salt) to BSH mixture ratio. The bar at the lower left corner of each image was 100 microns.
Fig. 13 is a graph showing the survival rate of cancer cells when neutron beam irradiates cancer cells including a complex containing A6K (TFA salt) and BSH. non B10 is a system in which neither complex nor BSH is added to the cell, BSH is a system in which only BSH is added to the cell, and BSH/A6K is a system in which a complex containing A6K (TFA salt) and BSH is added to the cell.
FIG. 14 is a view showing specific introduction of BSH into a tumor site by the drug of the present invention. The upper panel (indicated with only BSH) shows the case of BSH administration alone, and the lower panel (A6K/BSH (compound described in example 4 (1)) shows tumor tissue when the drug of the present invention (complex containing A6K and BSH) is administered Hoechst shows a nuclear staining pattern, HLA-a shows an HLA-a immunostaining pattern, BSH shows a BSH immunostaining pattern, and Merge shows a pattern obtained by staining three layers on the same screen.
Detailed Description
In one embodiment, the present invention provides a method of preparing a complex comprising a peptide comprising a hydrophobic amino acid residue and a basic amino acid residue and BSH, the method comprising mixing the peptide and BSH in an aqueous solution. Hydrophobic amino acids and basic amino acids are well known. Various kinds of peptides containing hydrophobic amino acid residues and basic amino acid residues are also known to be useful in the present invention. In the present invention, a peptide containing a hydrophobic amino acid residue and a basic amino acid residue, which is preferably used, is represented by the following formula (1):
(X)m-(Z)n (1)
wherein each of the m amino acid residues X is independently alanine, valine, leucine, or glycine; n amino acid residues Z are each independently-NHCH (COOH) R1；R1Is- (CH)2)pNHR2；R2is-H or-C (NH) NH2(ii) a m is 4 to 10; n is 1 to 2; and p is 1 to 6; in the present invention, more preferably used examples of the peptides containing hydrophobic amino acid residues and basic amino acid residues include XXXXZ, XXXXZZ, XXXXXXXXXZ, XXXXXXZZ, XXXXXXXZ, and XXXXXXXXZZ, but are not limited thereto. Specific examples of these peptides include AAAAAK, AAAAAAK, AAAAAAAK, AAAAAKK, AAAAAAAKK, AAAAAR, AAAAAAR, AAAAAAAR, AAAAARR, AAAAAARR, and AAAAAAARR. In the present invention, more preferably used containing hydrophobic amino acid residues and basic amino acid residues of peptides further specific examples include AAAAAA-high arginine, AAAAAA-ornithine, AAAAAA-2, 7-two amino acid, AAAA-2, 4-two amino acid, and AAAA-2-amino-4-guanidino butyric acid. Typical examples of the above-described peptides more preferably used in the present invention include AAAAAAK (abbreviated as A6K) and aaaaaaaar (abbreviated as A6R). Amino acid residues X and Z may be modified when a modifiable moiety is present in amino acid residues X and Z. Furthermore, 1 or 2 amino acid residues in the peptide represented by formula (1) may be substituted with amino acid residues other than hydrophobic amino acid residues and basic amino acid residues. In the method of making the complexes of the invention, the peptide may be in free form, salt form, solvate form, or modified or derivatized. Various salts of peptides are known, and methods for their preparation are also known. Examples of the salt of the peptide include hydrochloride, sulfate, nitrate, phosphate, acetate, trifluoroacetate (TFA salt), citrate, succinate, maleate, fumarate, malate, tartrate, p-toluenesulfonate, benzenesulfonate, methanesulfonate, alkali metal salt, and alkaline earth metal salt, but are not limited thereto. Solvates of the peptides are also well known and methods for their preparation are also well known. Examples of the solvate of the peptide include solvates of water, methanol, ethanol, isopropanol, THF, DMSO, ethylene glycol, propylene glycol, and acetamide, but are not limited thereto. Various modified peptides and peptide derivatives are known. Methods of modifying and derivatizing peptides are also well known. Examples of modifications and derivations of peptides include, but are not limited to, alkylation such as acetylation, amidation, biotinylation, maleimidation (maleimidation), and methylation, maleimidation, myristoylation, esterification, phosphorylation, and labeling such as fluorescent labeling and radioactive labeling, among others. The N-terminus of the peptide is preferably acetylated. Further, the amino acids constituting the peptide may be natural amino acids or unnatural amino acids, and may be in the L-form or D-form.In the present invention, when mentioned, the peptide includes modified peptides, derived peptides, salt form peptides, amino acid substituted peptides, and D amino acid-containing peptides as described above. The peptides listed above as examples are represented by the conventional one-letter amino acid code.
BSH is also well known and is a molecule having 12 boron isotopes in one molecule10B, a crystal of B. As described above, BSH has many boron atoms per molecule, and has high collision efficiency with neutron beams when used for BNCT. However, the killing effect of cancer cells by BNCT is low, since BSH cannot penetrate the cell membrane and thus does not enter the cell itself. In the present invention, a complex comprising a peptide containing hydrophobic amino acid residues and basic amino acid residues and BSH was constructed, thereby successfully delivering and retaining BSH in cancer cells. Thereby, the cancer cell killing effect by BNCT can be significantly enhanced and safety can be ensured at the same time. In the method of making the complexes of the invention, BSH may be modified or derivatized. Modified BSH and derivatized BSH are well known, and examples shown include peptide-bound BSH, saccharide-bound BSH, and BSH having thiol, hydroxyl, carboxyl, amino, amide, azide, halogen, and phosphate groups, but are not limited thereto. Methods for preparing modified and derivatized BSH are also well known.
The complex comprising the peptide containing the hydrophobic amino acid residue and the basic amino acid residue and the BSH of the present invention can be obtained by mixing the peptide and the BSH in an aqueous solution. This operation is very simple. When mixing, stirring or sonication may be performed if necessary. The concentration of the peptide in the aqueous solution is not particularly limited and is usually several μ M to several thousand μ M. The concentration of BSH in the aqueous solution is not particularly limited and is usually several tens μ M to several thousands μ M. The molar ratio of the above-mentioned peptide and BSH to be mixed is not particularly limited, but the mixing ratio is preferably a ratio of about 1mol to about 1000mol of BSH to 1mol of peptide, and for example, a ratio of about 1 to about 100mol of BSH to 1mol of peptide, and may be a ratio of about 100 to about 1000mol of BSH to 1mol of peptide.
The aqueous solution used for mixing to prepare the complex comprising the peptide containing the hydrophobic amino acid residue and the basic amino acid residue and BSH is a solution having water as a medium. The aqueous solution may be water alone, or other substances such as a buffer or salt may be added thereto. The conditions such as temperature, pH and mixing time at the time of mixing can be determined by one skilled in the art if desired. For example, when BSH is mixed with A6K or A6R, it is preferably mixed under the condition that the lysine residue of A6K or the arginine residue of A6R is positively charged and BSH is negatively charged. The pH of the aqueous solution can be adjusted to a desired value using a buffer such as PBS.
The complex comprising the peptide containing the hydrophobic amino acid residue and the basic amino acid residue and BSH obtained by the method of the present invention has a spherical shape having angular protrusions on the surface or a spherical shape without such protrusions. Spherical includes not only perfect spheres but also substantially spheres. Specifically, the shape is defined as a sphere when the ratio of the short diameter to the long diameter is about 0.5 or more, preferably about 0.6 or more, and further preferably about 0.7 or more. The diameter of the composite obtained by the present invention is about 20nm to about 200 nm. The diameter of the composite is an average of the major and minor diameters, and when the sphere has angular protrusions, the diameter includes the protrusions. The diameter of the complex of the present invention can be measured, for example, by using an electron microscope or the like. In the present specification, for example, when "the diameter of the composite is about 20nm to about 200 nm", this means that the majority of the composite, for example, about 50% or more, preferably about 60% or more, and further preferably about 70% or more, is about 20nm to about 200nm in diameter.
Among the various DDS carriers, the major factor associated with drug delivery is the size of the complex of drug and carrier. When the complex is too large in diameter, it is difficult to leak out of the tumor vessels, resulting in low tumor accessibility. When the diameter is too small, the drug retentivity at the tumor portion becomes low, resulting in a low drug concentration. The ideal value of the complex of drug and carrier reported so far is about 20nm to 100 nm. In other words, the diameter is about 100nm when it is expected that the EPR effect is produced by an increase in vascular permeability around a cancer tissue site, but in the case of refractory cancer accompanied by chronic inflammation, the diameter can be said to be about 20nm to 30 nm. As explained below, the complexes of the invention have a diameter of about 20nm to about 200nm, for example, about 20nm to 100nm, which is close to the ideal value, so that both tumor accessibility and drug retention are high.
According to the preparation method of the present invention, many complexes having a diameter of about 20nm to about 200nm can be simply obtained by mixing the peptide and the BSH. Therefore, the complex obtained by the preparation method of the present invention can be used for BNCT as such. Many complexes have small diameters when the ratio of BSH to peptide to be mixed is low, whereas many complexes have large diameters when the ratio of BSH to peptide is high. The diameter of the composite can be adjusted using this characteristic. Alternatively, the diameter of the composite may be adjusted using a filter having a desired pore size or an extruder having a desired pore size.
In another embodiment, the present invention provides a complex comprising a peptide comprising a hydrophobic amino acid residue and a basic amino acid residue and BSH. The composites of the present invention are spherical with a diameter of about 20nm to about 200 nm.
In yet another embodiment, the invention provides a medicament comprising the above complex for BNCT in cancer. The complexes of the invention can efficiently deliver and retain BSH in cancer cells. Therefore, the effect of BNCT can be obviously enhanced by the medicament containing the complex.
The cancer that can be treated using the drug of the present invention may be any kind of cancer, and is not particularly limited. Examples of cancers that can be treated using the drug of the present invention include esophageal cancer, gastric cancer, colorectal cancer, liver cancer, gallbladder cancer, bile duct cancer, pancreatic cancer, renal cell carcinoma, gastrointestinal stromal tumor, mesothelioma, brain tumor (meningioma, glioma, pituitary tumor, acoustic neuroma, glioblastoma multiforme, etc.), head and neck cancer, laryngeal cancer, oral cancer, cancer of the floor of the mouth (cancer of the mouth), gum cancer, tongue cancer, buccal mucosa cancer, salivary gland cancer, paranasal sinus cancer, maxillary sinus cancer (maxillary sinus cancer), frontal sinus cancer, ethmoid sinus cancer, sphenoid sinus cancer, thyroid cancer, lung cancer, osteosarcoma, bladder cancer, prostate cancer, testicular cancer, penile cancer, breast cancer, endometrial cancer, cervical cancer, ovarian cancer, skin cancer, rhabdomyosarcoma, leukemia, lymphoma, hodgkin's disease, non-hodgkin's lymphoma, and the like, And multiple myeloma, but not limited thereto.
The complexes of the invention can be used as such as a medicament for BNCT, or can be formulated into various dosage forms using pharmaceutically acceptable carriers or excipients by methods well known to those skilled in the art. The carriers or excipients used are well known to those skilled in the art and may be selected as desired. The medicaments of the invention may be prepared using means and methods well known to those skilled in the art. For example, when preparing injections and infusion solutions, pharmaceutically acceptable carriers such as physiological saline or phosphate buffered saline may be used. For the preparation of the medicament of the present invention, pharmaceutically acceptable additives such as thickening agents, absorption enhancers, pH adjusting agents, preservatives, dispersing agents, wetting agents, stabilizers, preservatives, suspending agents, and surfactants can be used.
The dosage form of the drug of the present invention is not particularly limited and may be selected, if necessary, according to the site, size, kind and patient state of the cancer to be treated. The drug of the present invention may be in a liquid, semi-solid, or solid state. Examples of the dosage form of the drug of the present invention include injections, infusion solutions, nasal drops, eye drops, lotions, sprays, ointments, gels, ointments, suppositories, tablets, capsules, powders, granules, syrups, aerosols, transdermal agents, transmucosal agents, and inhalants, but are not limited thereto. Alternatively, the drug of the present invention may be in the form of a lyophilized product, which is suspended in a pharmaceutically acceptable carrier such as physiological saline or phosphate buffered saline when administered.
The route of administration of the drug of the present invention is not particularly limited, but may be selected as necessary depending on the site, size, kind and patient condition of the cancer to be treated. Examples of routes of administration of the drug of the present invention include, but are not limited to, topical administration such as subcutaneous injection, intradermal injection, intravenous injection, infusion, oral administration, transmucosal administration, intestinal administration, eye drop administration, nasal administration, ear drop, inhalation, transdermal administration, and intratumoral administration, and intracerebroventricular administration.
The dosage of the drug of the present invention can be determined by the physician, if necessary, depending on the site, size, kind and patient condition of the cancer to be treated.
After the drug of the invention has been administered to the patient and a time sufficient for the complex of the invention to reach the site to be treated has elapsed, it is subsequently irradiated with a neutron beam. At the time of neutron irradiation, a reactor or an accelerator type neutron generator is used, and conditions necessary for treatment, such as a neutron beam dose and a neutron spectrum, and irradiation time, are determined.
Administration of the drug of the present invention and neutron irradiation may be performed once to many times. The physician may determine the number of times in consideration of the location and type of cancer, the degree of reduction in the size of cancer, the patient's condition, and the like.
In another embodiment, the present invention provides the use of the above complex for the preparation of a medicament for BNCT in cancer.
In another embodiment, the present invention provides the use of the above complex for BNCT in cancer.
In another embodiment, the invention provides a method of cancer treatment comprising administering the above complex to a cancer patient and irradiating the patient with neutrons.
Hereinafter, the present invention is described in further detail specifically with reference to examples, but the examples should not be construed as limiting the scope of the present invention.
[ example 1]
(1) Shape of A6K (TFA salt) in aqueous solution
The lyophilized product of A6K (TFA salt) synthesized by conventional methods was dissolved in Milli-Q water (concentration 1000. mu.M) and the pH was adjusted to 4 with HCl. Sonication was carried out for 10 minutes, adjusting the pH to 7 with NaOH. The resulting solution was passed through an extruder having a pore size of 100nm and diluted with Milli-Q water to a concentration of 50. mu.M. In this solution, 1.2. mu.L was divided into samples and observed using a scanning electron microscope. A scanning electron micrograph is shown in figure 1. A6K (TFA salt) was found to be tubular. A DLS assay was performed on a solution of A6K (TFA salt) at a concentration of 200 μ M prepared in a similar manner as described above. A graph is shown in fig. 2. In the graph, a bimodal peak was observed, demonstrating that A6K (TFA salt) is tubular.
(2) Production of A6K (TFA salt) and BSH-containing Complex
An aqueous solution (10. mu.M) of A6K (TFA salt) in Milli-Q water was obtained in a similar manner to (1) above. BSH (concentration 1000 μ M) was added and mixed with stirring at room temperature for 3 minutes, to observe the resulting complex using a scanning electron microscope. The results are shown in fig. 3. The complexes were found to be spherical with horny protrusions (gold candy shapes) and most of them had diameters ranging from about 20nm to about 150 nm. The DLS test was carried out on a complex obtained by mixing A6K (TFA salt) (200. mu.M) and BSH (2000. mu.M) in a similar manner to that described above. The results are shown in FIG. 4. Bimodality peaks were observed adjacent to each other, demonstrating that the complex was roughly spherical. These results show that the shape and size of the resulting complex is optimal for delivering BSH into cancer cells.
(3) Delivery of complexes into cancer cells
The complex was prepared by mixing A6K (TFA salt) and BSH with stirring in the same manner as described in (2) above. The mixing time was 10 minutes, 30 minutes, and 180 minutes. The resulting complexes were diluted with PBS (pH 7.1 to 7.3) and added to the glioma cell line U87 Δ EGFR in culture dishes. The complex was added at a final concentration of 50 μ M A6K (TFA salt) and 5000 μ M BSH. ICP-AES was performed on each cell sample obtained by culturing the complex at 37 ℃ for 3 hours, 12 hours, and 24 hours, thereby determining the boron concentration in the cells. The results are shown in FIG. 5. The complex enters the cell under all conditions. It has been found that large amounts of complex enter the cells in 12 and 24 hours of culture. A12 hour incubation time is sufficient. The mixing time with stirring had little effect on the amount of complex delivered to the cancer cells. A stirring and mixing time of 10 minutes is sufficient. As the culture time was extended, an increase in the amount of BSH in the cells was detected. Thus, the complex has been demonstrated to have retention in cancer cells.
[ example 2]
(1) Production of A6R (TFA salt) and BSH-containing Complex
The lyophilized product of A6R (TFA salt) synthesized by conventional methods was dissolved in Milli-Q water (concentration 1000. mu.M) and the pH was adjusted to 4 with HCl. Sonication was carried out for 10 minutes, adjusting the pH to 7 with NaOH. The resulting solution was diluted with Milli-Q water to a concentration of 10. mu.M. BSH was added to the thus obtained aqueous solution (10 μ M) of A6R (TFA salt) in Milli-Q (concentration 1000 μ M), mixed with stirring at room temperature for 3 minutes and subjected to sonication for 10 minutes, followed by passing through an extruder having a pore size of 50nm, to thereby obtain a complex. The resulting complex was observed using a scanning electron microscope. The results are shown in FIG. 6. It has been found that the composites are spherical and that most of them have a diameter in the range of about 100nm to about 200 nm. Further, complexes were prepared in a similar manner as described above except that the molar ratio of A6R (TFA salt) to BSH was 1:1 (20. mu.M: 20. mu.M), the size of the complexes was smaller, with a majority of them having diameters of about 50nm to about 150nm, including those having diameters of about 20 nm.
(2) Delivery of complexes into cancer cells
The complex was prepared by mixing A6R (TFA salt) and BSH with stirring in the same manner as described in (1) above. The resulting complexes were diluted with PBS (pH 7.1 to 7.3) and added to the glioma cell line U87 Δ EGFR in culture dishes. The complex was added in an amount such that the final concentrations of A6R (TFA salt) and BSH were as shown in fig. 7 and 8. ICP-AES was performed on each cell sample obtained by incubation with the complex at 37 ℃ for 6 hours, 12 hours, and 24 hours, to thereby determine the boron concentration in the cells. The results are shown in FIGS. 7 and 8. As shown in fig. 7, it was confirmed that a complex of A6R (TFA salt) and BSH was introduced into cells in a concentration-dependent manner. Further, as shown in FIG. 8, a culture time of 6 hours is sufficient. After 12 hours and 24 hours of culture, a considerable amount of the complex remained in the cells, demonstrating that the complex was retained in cancer cells.
[ example 3]
The intracellular distribution of the complexes of the invention was investigated.
(1) Experimental methods
In a 24-well plate (manufactured by Falcon) on a glass plate (PLL-coated, 12 mm: IWAKI)&CO., ltd. manufacture) U87 Δ EFGR cells (3000 cells/well, 1ml in each well) were cultured and CO at 37 ℃ in air2Cultured in an incubator 24In the course of hours, a complex containing A6K (TFA salt) and BSH, prepared in the same manner as in example 1(2), was added. The complex was added in an amount such that the final concentration of A6K (TFA salt) was 20. mu.M and the final concentration of BSH was 2000. mu.M. After addition of the complex, the cells were cultured for 90 minutes, the cell culture solution was removed, 1ml of PBS (phosphate buffered saline) was added at room temperature, the cells were allowed to stand for 5 minutes, and thereafter removed and washed 3 times (5 minutes for 1ml each). Then, Paraformaldehyde (PFA) solution (4%, 1ml) was added, and the cells were cultured for 30 minutes and fixed. Cells were washed 3 times with PBS (supra). Thereafter, a PBS solution (1ml) containing Triton (0.25%) was added and the cells were cultured at 37 ℃ for 15 minutes. Cells were washed 3 times with PBS (supra). Then, a PBS solution (1ml) containing BSA (bovine serum albumin, 1%) was added and the cells were cultured at room temperature for 1 hour. Thereafter, the cells were washed 3 times with PBS (same as above).
A sufficient amount of primary antibody staining solution (BSH antibody [1:200] (final concentration 0.5. mu.g/ml) in 0.1% BSA PBS) was added to cover the samples except for the negative control. For the negative control, a solution without primary antibody for diluting the antibody was used. Samples were incubated for 2 hours at room temperature, respectively. The antibody staining solution was removed from the sample once, after which the cells were washed 3 times with PBS (same as above). A sufficient amount of secondary antibody staining solution (donkey anti-mouse IgG in 0.1% BSA PBS (Alexa 488) [1:100]) was added to cover the samples, which were incubated at room temperature for 2 hours. The same procedure was performed for a negative control containing no secondary antibody. The secondary antibody staining solution was removed from the sample, after which the cells were washed 3 times with PBS (same as above).
A potting tablet (mount) (ProLong (registered trademark) Diamond: Thermo) was added to the glass preparation to fix the sample. Thus, a cellular immunostaining glass preparation of cancer cells U87 Δ EGFR was prepared.
(2) Results of the experiment
A stained image is shown in fig. 9. Staining was detected not only in the cytoplasm but also in the nucleus, thus demonstrating that the complex of the invention moves not only into the cell but also into the nucleus. From these results, it can be said that the cells containing the complex of the present invention are irradiated with a neutron beam to selectively destroy the cells, thereby achieving effective cancer treatment.
[ example 4]
The inhibition of colony formation was investigated by neutron irradiation of cancer cells containing the complex of the invention.
(1) Production of complexes containing A6K (hydrochloride salt) and BSH
The lyophilized product of A6K (hydrochloride salt) synthesized by a conventional method was dissolved in Milli-Q water and sonicated for 10 minutes. A portion of the resulting aqueous solution of A6K (hydrochloride salt) in Milli-Q was isolated and observed with scanning electron microscopy and transmission electron microscopy. A6K (hydrochloride salt) has a tubular shape. BSH was added to an aqueous solution of A6K (hydrochloride salt) in Milli-Q, mixed with stirring at room temperature for 3 minutes, and sonicated for 10 minutes. Spherical complexes were obtained when the molar ratio of A6K (hydrochloride salt) to BSH was 1:10 (166. mu.M: 1.66mM) and 1:25 (166. mu.M: 4.15 mM). The majority of the resulting composite had a diameter of about 100 nm. A scanning electron micrograph of the complex when the molar ratio of A6K (hydrochloride salt) to BSH was 1 to 10 is shown in fig. 10.
(2) Delivery of complexes into cancer cells
The complex containing A6K (hydrochloride) and BSH obtained in (1) above (A6K (hydrochloride): BSH molar ratio 1:10) was added to the glioma cell line U87 Δ EGFR in a similar procedure as described in example 2 (2). The complex was added in an amount such that the final concentrations of A6K (hydrochloride salt) and BSH were as shown in fig. 11. The cells and the complex were cultured at 37 ℃ for 24 hours, followed by ICP-AES to determine the boron concentration in the cells. As shown in FIG. 11, the group with only BSH (2mM) addition had 595.2. + -. 105.1ng/106Intracellular boron concentration of individual cells, whereas the group to which the complex of A6K (hydrochloride) (200. mu.M) and BSH (2mM) was added had 11262. + -. 3890ng/106Intracellular boron concentration of individual cells. From these results, it was found that the present complex specifically entered cancer cells and a high concentration of the complex remained in the cells even after 24 hours of culture.
(3) Localization of intracellular complexes
The intracellular distribution of the complex of A6K (hydrochloride salt) and BSH delivered into the cells was investigated in a similar procedure as described in example 3 (1). A micrograph of cancer cells stained with BSH-specific antibodies is shown in fig. 12. According to these results, intracellular introduction of BSH as a boron drug was confirmed in both the A6K (hydrochloride salt)/BSH complex 40. mu.M/400. mu.M administration group and the 200. mu.M/400. mu.M administration group. It can be said that the cells containing the complex of the present invention are selectively destroyed by neutron beam irradiation, thereby achieving effective cancer treatment.
[ example 5]
The cancer cells containing the complex of the present invention are irradiated with neutrons to perform a colony formation test.
The complex containing A6K (hydrochloride salt) and BSH obtained in example 4(1) was added to a cell line of squamous cell carcinoma origin (SAS) of the tongue in a similar procedure as described in example 2 (2). The complex was added in an amount such that A6K (hydrochloride salt) was 20. mu.M and BSH was 2000. mu.M (0.24 mg/ml based on B10). Tests were carried out on a system with only BSH (2000. mu.M) added (0.24 mg/ml as B10) and on a system without complex or BSH. Both systems were incubated at 37 ℃ for 24 hours. Neutron beam irradiation was performed for 20 minutes, 45 minutes, and 90 minutes. Cell viability was calculated by colony formation method. In each system, the number of samples (N) was 3. The results are shown in fig. 13. The survival of cancer cells comprising a complex comprising A6K (hydrochloride salt) and BSH has been shown to decrease significantly with increasing neutron fluence (neutron flux). When the neutron fluence is 6.7x1011n/cm2When the survival rate of cancer cells containing neither the complex nor BSH and the survival rate of cancer cells containing only BSH were 10 to 20%, the survival rate of cancer cells containing the above complex was only 2% or less. From these results, it was shown that when cells containing the complex of the present invention were irradiated with a neutron beam, such cells could be selectively destroyed, thereby proving that effective cancer treatment could be achieved.
[ example 6]
The drugs of the present invention were administered to brain tumor model animals (in vivo models implanted with U87 Δ EGFR) to investigate whether BSH was tumor-specifically introduced.
According to the procedures approved by Animal Care and Use Committee (Okayama University) of Okayama UniversityAnimals used in this experiment were bred, kept and used in the procedures (approved code: OKU-2016475). By mixing 3. mu.L of U87. delta. EGFR cell suspension (1X 10)5Cells/μ L) was injected directly into the brain to generate tumor-bearing model mice (BALB/C nu/nu, female, 6 to 8 weeks old, 25g, Japan SLC, inc., Shizuoka). After two weeks, 200. mu.L of the complex containing A6K and BSH (A6K 2mM/BSH 20mM), and 200. mu.L of BSH 20mM as a control experiment were administered separately from the tail vein (A6K. HCl was converted to 8mg/kg and BSH was converted to 33.4mg/kg, considering that the body weight of the mouse was 20 g).
The complex used in this experiment (compound described in example 4 (1)) was prepared as follows. 70 μ L of MQ water was added to 70 μ L of A6K 10mM to adjust to A6K 5 mM. A solution of A6K 2mM/BSH 20mM was prepared by adding 35. mu.L of BSH 200mM and 175. mu.L of MQ water to 140. mu.L of A6K 5 mM. A6K was obtained from 3-D Matrix, ltd, and BSH was obtained from STELLA PHARMA CORPORATION.
After 12 hours of administration, mouse brains were embedded with the embedding medium Tissue Tek o.c.t compound for frozen Tissue sections. After freezing, the brain was thinly sliced to a thickness of 10 μm to prepare frozen sections. Frozen sections were fixed at room temperature using 4% (w/v) paraformaldehyde (PFA, Wako Pure Chemical Corporation). After washing with PBS, sections were blocked with 1% (w/v) BSA and submerged in a solution containing 0.3% (v/v) TritonX-100 (5. mu.g/mL) of anti-BSH mouse monoclonal antibody and rabbit anti-HLA-A antibody for 2 hours at room temperature. After washing, the sections were immersed for 2 hours at room temperature in a solution (20. mu.g/mL) of donkey anti-mouse antibody (Life Technologies) labeled with Alexa-Fluor 488 (green fluorescence) and a solution (2. mu.g/mL) of goat anti-rabbit antibody (Life Technologies) labeled with Alexa-Fluor 555 (red fluorescence). After washing, the cell nuclei were stained with Hoechst 33258 to produce a preparation, which was then observed for the local presence of BSH using a laser confocal microscope. anti-BSH antibodies provided by professor Kirihata of Osaka Prefecture University were diluted and frozen and thawed at the time of use.
HLA-a is a human major histocompatibility antigen and is expressed in transplanted and expanded human-derived glioblastoma U87 Δ EGFR, while it is not expressed in normal brain tissue of mice. In this experiment, red fluorescence was used to detect HLA-A. It was demonstrated that human-derived glioblastoma U87 Δ EGFR was amplified in nude mouse brain due to the presence of cell populations with red fluorescence and clear borders, resulting in a human brain tumor model.
The compound described in example 4(1) (A6K/BSH) was administered into these brain tumor model animals in a single dose from the tail vein, and after 12 hours, the brain tumor tissue was removed, immunostained using an anti-BSH mouse monoclonal antibody, and the presence of BSH was detected using green fluorescence. For the control experiment, the same amount of BSH was administered in a single dose. A summary of the results is shown in figure 14.
A strong green color indicating the presence of BSH was detected in mice administered with the compound described in example 4(1) (A6K/BSH), and further, the green color was consistent with HLA-a positive (red) tumor tissue. The green color was weak in the control experiment with only BSH administration, thereby demonstrating that BSH did not enter cells in large amounts, and that the locations where BSH was present were not different between tumor tissue and normal brain tissue of mice.
In this experiment, a brain tumor model animal was produced by transplanting a human-derived glioblastoma into the brain of a nude mouse, and it was demonstrated that BSH can be site-specifically introduced by administering the compound described in example 4 (1).
Industrial applicability
The invention can be used for preparing cancer treatment medicines, in particular to the field of cancer radiotherapy.
Claims (10)
1. A method of making a composite body comprising a peptide comprising a hydrophobic amino acid residue and a basic amino acid residue, and mercaptoundehydroceledodecaborate (BSH),
the method comprises mixing the peptide with BSH in an aqueous solution,
wherein BSH is mixed in a ratio of 1mol to 1000mol with respect to 1mol of the peptide;
the composite is spherical with a diameter of about 20nm to about 200 nm;
the peptide is represented by the following formula (1):
(X)m–(Z)n (1)
wherein X is alanine; m is 6; z is lysine or arginine; and n is 1.
2. A complex comprising a peptide comprising hydrophobic and basic amino acid residues and BSH, wherein the complex is spherical with a diameter of about 20nm to about 200 nm;
the peptide is represented by the following formula (1):
(X)m–(Z)n (1)
wherein X is alanine; m is 6; z is lysine or arginine; and n is 1.
3. A medicament for boron neutron capture therapy of cancer comprising the complex of claim 2.
4. Use of the complex of claim 2 in the preparation of a medicament for treating cancer by a method comprising administering the complex of claim 2 to a cancer patient and irradiating the cancer patient with a neutron beam.
5. The complex of claim 2 for use in boron neutron capture therapy of cancer.
6. Use of the complex of claim 2 in the manufacture of a medicament for boron neutron capture therapy of cancer.
7. The medicament of claim 3, which is an intravenous or infusion solution.
8. The use according to claim 4, wherein the complex is administered by intravenous injection or infusion.
9. The complex of claim 5, which is administered by intravenous injection or infusion.
10. The use of claim 6, wherein the medicament is administered by intravenous injection or infusion.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2016-229302 | 2016-11-25 | ||
| JP2016229302 | 2016-11-25 | ||
| PCT/JP2017/043220 WO2018097335A1 (en) | 2016-11-25 | 2017-11-24 | Novel bsh complex for boron neutron capture therapy |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110198740A CN110198740A (en) | 2019-09-03 |
| CN110198740B true CN110198740B (en) | 2022-03-04 |
Family
ID=62196269
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201780084294.2A Active CN110198740B (en) | 2016-11-25 | 2017-11-24 | Novel BSH complexes for boron neutron capture therapy |
Country Status (5)
| Country | Link |
|---|---|
| US (2) | US10894088B2 (en) |
| EP (1) | EP3545961A4 (en) |
| JP (1) | JP6548060B2 (en) |
| CN (1) | CN110198740B (en) |
| WO (1) | WO2018097335A1 (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117085140A (en) | 2017-12-15 | 2023-11-21 | 立美基股份有限公司 | Surfactant peptide nanostructures and their use in drug delivery |
| JP7221539B2 (en) * | 2018-02-15 | 2023-02-14 | 国立大学法人 岡山大学 | Cancer lesion tissue evaluation to optimize the effect of boron neutron capture therapy |
| JP2023085151A (en) * | 2021-12-08 | 2023-06-20 | 株式会社八神製作所 | Stem cell fractionation method using in vitro boron neutron reaction |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008308440A (en) * | 2007-06-14 | 2008-12-25 | Genomidea Inc | Pharmaceutical delivery vehicle for cancer treatment, method for producing the same, and preparation using the same |
| JP5538224B2 (en) * | 2008-07-24 | 2014-07-02 | ステラファーマ株式会社 | Optically active α-amino acids into which BSH has been introduced and method for synthesizing the same |
| SG193822A1 (en) * | 2008-08-27 | 2013-10-30 | 3 D Matrix Ltd | Transfection agent |
| JP6009154B2 (en) * | 2011-10-19 | 2016-10-19 | 国立大学法人 岡山大学 | Cell membrane permeable boron peptide |
| WO2017164334A1 (en) * | 2016-03-23 | 2017-09-28 | 公益財団法人 川崎市産業振興財団 | Boron cluster bonded peptide compound |
-
2017
- 2017-11-24 US US16/463,747 patent/US10894088B2/en active Active
- 2017-11-24 WO PCT/JP2017/043220 patent/WO2018097335A1/en unknown
- 2017-11-24 JP JP2018553029A patent/JP6548060B2/en active Active
- 2017-11-24 EP EP17873605.4A patent/EP3545961A4/en active Pending
- 2017-11-24 CN CN201780084294.2A patent/CN110198740B/en active Active
-
2020
- 2020-09-10 US US17/017,171 patent/US10940210B2/en active Active
Non-Patent Citations (2)
| Title |
|---|
| "Lipid-like Self-Assembling Peptide Nanovesicles for Drug Delivery";Dimitrios G. Fatouros等;《ACS Appl. Mater. Interfaces》;20140512(第6期);第8184-8189页 * |
| "Synthesis and evaluation of cyclic RGD-boron cluster conjugates to develop tumor-selective boron carriers for boron neutron capture therapy";Sadaaki Kimura等;《Bioorganic & Medicinal Chemistry》;20110118(第19期);摘要，Figure1，Scheme1，第1726页左栏 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110198740A (en) | 2019-09-03 |
| EP3545961A4 (en) | 2020-07-29 |
| WO2018097335A1 (en) | 2018-05-31 |
| JPWO2018097335A1 (en) | 2019-04-18 |
| US10940210B2 (en) | 2021-03-09 |
| US10894088B2 (en) | 2021-01-19 |
| US20200121800A1 (en) | 2020-04-23 |
| JP6548060B2 (en) | 2019-07-24 |
| EP3545961A1 (en) | 2019-10-02 |
| US20200397910A1 (en) | 2020-12-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11406597B2 (en) | Fusogenic liposome-coated porous silicon nanoparticles | |
| US20210212948A1 (en) | Nanovesicles derived from cell membrane, and use thereof | |
| US10940210B2 (en) | BSH complex for boron neutron capture therapy | |
| EP2983720B1 (en) | Platform for targeted delivery to tumor cells and uses thereof | |
| US11337924B2 (en) | Targeted polymerized nanoparticles for cancer treatment | |
| CN105163721A (en) | Lipid nanoparticle compositions and methods of making and methods of using the same | |
| Michiue et al. | Self-assembling A6K peptide nanotubes as a mercaptoundecahydrododecaborate (BSH) delivery system for boron neutron capture therapy (BNCT) | |
| CN107875384A (en) | A kind of neoplasm targeted therapy drug delivery system for containing sensitising agent | |
| Li et al. | A hybrid of lactic acid bacteria and metal-organic frameworks potentiates photodynamic immunotherapy for hypoxia-targeted tumor eradication | |
| Li et al. | Ginsenoside Rg3-engineered exosomes as effective delivery platform for potentiated chemotherapy and photoimmunotherapy of glioblastoma | |
| CN113271928A (en) | Nanocarriers for pulmonary inflammation treatment | |
| Li et al. | Lipid Nanocarrier‐Based mRNA Therapy: Challenges and Promise for Clinical Transformation | |
| US9526791B2 (en) | Weakly acidic pH-responsive peptide and liposome containing same | |
| Guo et al. | β-glucan-modified nanoparticles with different particle sizes exhibit different lymphatic targeting efficiencies and adjuvant effects | |
| JP2022530539A (en) | Nanostructures containing cobalt porphyrin-phospholipid conjugates and polyhistidine tags | |
| US9944676B2 (en) | Cationic lipid formulations for regressing established tumor | |
| Suzuki | Project 10 Preclinical Study for Development of New Drug for NCT (29P10) | |
| Wang et al. | Amifostine loaded lipid-calcium carbonate nanoparticles as an oral drug delivery system for radiation protection | |
| CN118141765A (en) | FAP activatable liposome capable of targeted delivery of ATRA and preparation method and application thereof | |
| Zhang et al. | Hyaluronate-based Self-stabilized Nanoparticles for Reverting Immunosuppression and Immunochemotherapy in Osteosarcoma Treatment | |
| Hu | Bioinspired Drug Delivery System for Anticancer Therapy | |
| EP4408471A2 (en) | Borylated di-peptide amino acid compositions for use in boron neutron capture therapy and methods thereof | |
| CN118480088A (en) | Cationic polypeptide capable of killing cancer cells in inflammatory manner and application thereof | |
| Wang | Polymer-prodrug conjugate-based nanovesicles for immunoactivation | |
| Thallaj | Advancements in Peptide Vectors for Cancer Therapy and Tumor Imaging: A Comprehensive |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |